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A new growth factor indispensable for the growth of " True Hiochi bacteria " ) which were recently named as Lact. homochiochi and Lact. heterohiochi2> was isolated and purified. The Sake (Japanese rice wine) which is frequently spoiled by the " Hiochi " bacteria, contains this factor. It was not detected in liver extract, yeast extract, tomato juice, whey and laboratory peptones, but was found to be considerably accumulated in the culturebroth of several organisms, such as Aspergillus, Penicillium, Monilia, Willia and Lactobacillus. By means of bioautography it was confirmed that the " Hiochi " factors produced by these organisms in the broth were all the same;. From the broth in which Aspergillus oryzae was grown, the " Hiochi " factor was extracted with butanol and/or ethylacetate after being acidified to pH 2.0. A highly purified barium salt of the factor was obtained by the chromatographic technique using silica ge14'5). When the barium salt was acetylated with acetyl chloride or acetic anhydride, its activity for growth disappeared completely. However, the activity was recovered after alkalihydrolysis of the acetyl derivative, a fact indicating the presence of hydroxyl group. The p-bromophenacylester was not obtained in crystalline form, whereas crystallization was succeeded for the quinine salt and the S-benzylthiuromium salt (Fig. 1). Analyses of these two salts gave the following results. Quinine salt; m.p. 137-8°C. Anal. Calculated for Cz~H3606N2: C, 66.08; H, 7.65; N, 5,93. Found: C, 66.01,66.17; H, 7.64, 7.51; N, 5.92. S-benzylthiuronium salt; m.p. 122-3°C. Anal. Calculated for C,4H~~O4N~S: C, 53.49; H, 7.05; N, 8.91; S, 10.18, Found: C, 53,13, 53.41; H, 7.11, 7.16; N, 9.05; S, 9.49. The " Hiochi " factor freed from its quinine salt was then distilled in vacuo. A viscous syrupy material obtained showed the following charac-

HIOCHIC ACID, A NEW GROWTH FACTOR FOR LACTOBACILLUS HOMOHIOCHI AND LACTOBACILLUS HETEROHIOCHI